Contents

Overview

Dot Blot is a kind of Western Blot (without having to run a gel and doing a transfer). It is mostly used to determine the antibody titre, before performing the actual Western Blot. Dot Blot helps to determine the amount of primary or the secondary antibody that is needed to run a CLEAN Western Blot.

Materials

PVDF membrane

Wash Buffer

Blocking agent

Primary antibody

Secondary antibody

Detection reagent (ECL)

Procedure

Place 10-20μL of protein solution (diluted using sample loading buffer) on the membrane.